Stabilization of a reaction intermediate as a catalytic device: definition of the functional role of the flexible loop in triosephosphate isomerase

The function of the mobile loop of triosephosphate isomerase has been investigated by deleting four contiguous residues from the part of this loop that interacts directly with the bound substrate. From the crystal structure of the wild-type enzyme, it appears that this excision will not significantly alter the conformation of the rest of the main chain of the protein. The specific catalytic activity of the purified mutant enzyme is nearly 10(5)-fold lower than that of the wild type. Kinetic measurements and isotopic partitioning studies show that the decrease in activity is due to much higher activation barriers for the enolization of enzyme-bound substrate. Although the substrates bind somewhat more weakly to the mutant enzyme than to the wild type, the intermediate analogue phosphoglycolohydroxamate binds much less well (by 200-fold) to the mutant. It seems that the deleted residues of the loop contribute critically to the stabilization of the enediol phosphate intermediate. Consistent with this view, the mutant enzyme can no longer prevent the loss of the enediol phosphate from the active site and its rapid decomposition to methylglyoxal and inorganic phosphate. Indeed, when glyceraldehyde 3-phosphate is the substrate, the enediol phosphate intermediate is lost (and decomposes) 5.5 times faster than it reprotonates to form the product dihydroxyacetone phosphate.(ABSTRACT TRUNCATED AT 250 WORDS)     

Cyanobacterial metabolites as a source of sunscreens and moisturizers: a comparison with current synthetic compounds

The recognition that ultraviolet radiation has harmful effects on the skin has led to the commercial development of inorganic and synthetic organic UV filters that can reduce the negative effects of exposure to sunlight. In addition, moisturizing chemicals are extensively used in personal care products to improve the ability of skin to retain water. Whilst current UV filter and moisturizing chemicals have clear beneficial qualities, they may also have adverse effects such as contact sensitivity, oestrogenicity and even tumorigenic effects on human skin. Furthermore, the accumulation of these chemicals in the aquatic environment could be potentially harmful. Consequently, there is interest in exploiting safer alternatives derived from biological sources, especially from photosynthetic organisms such as cyanobacteria which have developed mechanisms for coping with high UV irradiation and desiccation. In order to overcome the detrimental effects of UV radiation, these microorganisms produce UV screening compounds such as mycosporine-like amino acids and scytonemin, which are good candidates as alternatives to current synthetic UV filters. In addition, extracellular substances produced by some extremophilic species living in hyper-arid habitats have a high water retention capacity and could be used in cosmetic products as moisturizers. In this review, we present an overview of the literature describing the potential of cyanobacterial metabolites as an alternative source for sunscreens and moisturizers.     

Small molecule of sphingosine as a rescue of dopaminergic cells: A cell therapy approach in neurodegenerative diseases therapeutics

Multiple sclerosis (MS) patients should take medication such as fingolimod (FTY-720) for a long time, hence pharmaceutical effects on other neural cells such as dopaminergic cells are important. Dopaminergic cell line, BE(2)-M17, was treated by FTY-720 and then cell viability and genes involve in neurosurvival were investigated. It was disclosed that FTY-720 significantly stimulates Bcl2 overexpression. Whereas, it decreased intracellular reactive oxygen species production and cell membrane damage of dopaminergic cells. The increase in Bcl2/Bax ratio increased the cell metabolic activity and decreased propidium iodide-positive cells. Besides, FTY-720 induced the overexpression of CACNA1C, nNOS gene, and nitric oxide production. However, FTY-720 induced GABARA1 overexpression and eventually it could overcame to the cytotoxic effect of intracellular calcium. This cascade led to tyrosine hydroxylase and BDNF genes overexpression whereas FTY-720 did not change GDNF concentration in BE(2)-M17 cells. Concluding, it might be said that taking FTY-720 in MS patients did not induce adverse effect on dopaminergic cells.     

Plant growth promoting activity of an auxin and siderophore producing isolate of <Emphasis Type="Italic">Streptomyces</Emphasis> under saline soil conditions

A biocontrol Streptomyces isolate (C) was tested for its plant growth promoting qualities under saline conditions. Exposure to elevated osmotic strengths up to 300 mM NaCl increased dry weight and cfu/ml significantly. The isolate C produced indolyl-3-acetic acid (IAA) into the medium in the amount of 2.4 μg/ml. The amount of auxin increased after adding salt and reached to 4.7 μg/ml in 300 mM NaCl. Biosynthesis of siderophore was detectable and increased in presence of NaCl. Streptomyces isolate C showed good solubilization of tricalcium phosphate in culture medium with 92 mg/l. Solubilization decreased in presence of NaCl. Soil treatment with isolate C increased the growth and development of wheat plant in normal and saline conditions. In this treatment there were significant increases in germination rate, percentage and uniformity, shoot length and dry weight compared to the control. Applying the bacterial inocula increased the concentration of N, P, Fe and Mn in wheat shoots grown in normal and saline soil, but had non-significant effect on other micro and macronutrients concentrations. Results of this study show that Streptomyces isolate C has potential to be utilized as biofertilizer in saline soils.     

Antimicrobial activity and chemical composition of the essential oil of Nepeta crispa Willd. from Iran

The composition and antimicrobial activity of the essential oil of Nepeta crispa Willd., an endemic species from Iran, was studied. The oil was obtained from the aerial parts of the plant and analyzed by GC and GC/MS. Twenty-three compounds, accounting for 99.8% of the total oil, were identified. The main constituents were 1,8-cineol (47.9%) and 4aalpha,7alpha,7abetanepetalactone (20.3%). The antimicrobial activity of essential oil of N. crispa was tested against seven gram-negative or gram-positive bacteria and four fungi. The results of the bioassays showed the interesting antimicrobial activity, in which the gram-positive bacteria, Bacillus subtilis and Staphylococcus aureus, were the most sensitive to the oil. Also, the oil exhibited a remarkable antifungal activity against all the tested fungi.     

Bacterial infections in the early period after liver transplantation in adults: A prospective single-center cohort study

Liver transplantation (LT) is a potentially curative treatment for terminal stage hepatic diseases. Bacterial infections are the main causes of mortality and morbidity in the early period after LT. Identifying the risk factors could help in minimizing their development. We prospectively investigated the incidence, characteristics, and risk factors of bacterial infections among the recipients during hospitalization after LT and assigned a predictive score. All 389 consecutive adults who underwent LT at the main referral hospital of LT in Iran during 1 year were enrolled prospectively in a cohort study. Infection group consisted of 143 recipients (36.8%). Urinary tract and surgical site infections were the most frequent ones. Gram-negative bacteria were more prevalent than Gram-positive ones. Independent risk factors were female sex (relative risks = 2.13), age ≤ 43.5 years (3.70), hospital stay ≥ 9.5 days (5.22), abdominal reoperation (3.03), vancomycin-resistant Enterococci colonization (5.52), hospitalization 3 months prior to LT (3.25), mechanical ventilation ≥48 hr (4.93), and renal replacement therapies (13.40). We developed a risk score for the prediction of bacterial infections with an area under the receiver operating characteristic curve of 0.85 (95% CI, 0.81–0.89) with sensitivity of 88% and specificity of 64%. In the infection group, mortality was higher than in controls (18.9% vs. 2.0%) with longer hospitalization (16 vs. 10 days; P < 0.001). We detected a high rate of bacterial infections leading to longer hospital stay and higher mortality rate. The formulated risk score can help predict bacterial infections; however, it requires clinical validation in further studies.     

Myeloid deletion and therapeutic activation of AMPK do not alter atherosclerosis in male or female mice

The dysregulation of myeloid-derived cell metabolism can drive atherosclerosis. AMP-activated protein kinase (AMPK) controls various aspects of macrophage dynamics and lipid homeostasis, which are important during atherogenesis. Using LysM-Cre to drive the deletion of both the α1 and α2 catalytic subunits (MacKO), we aimed to clarify the role of myeloid-specific AMPK signaling in male and female mice made acutely atherosclerotic by injection of AAV vector encoding a gain-of-function mutant PCSK9 (PCSK9-AAV) and WD feeding. After 6 weeks of WD feeding, mice received a daily injection of either the AMPK activator A-769662 or a vehicle control for an additional 6 weeks. Following this (12 weeks total), we assessed myeloid cell populations and differences between genotype or sex were not observed. Similarly, aortic sinus plaque size, lipid staining, and necrotic area did not differ in male and female MacKO mice compared with their littermate floxed controls. Moreover, therapeutic intervention with A-769662 showed no treatment effect. There were also no observable differences in the amount of circulating total cholesterol or triglyceride, and only minor differences in the levels of inflammatory cytokines between groups. Finally, CD68+ area and markers of autophagy showed no effect of either lacking AMPK signaling or AMPK activation. Our data suggest that while defined roles for each catalytic AMPK subunit have been identified, complete deletion of myeloid AMPK signaling does not significantly impact atherosclerosis. Additionally, these findings suggest that intervention with the first-generation AMPK activator A-769662 is not able to stem the progression of atherosclerosis.     

Evaluation of Exposure to Radiofrequency Electromagnetic Fields from Smart Utility Meters operating at 868 MHz

Power density and duty factor values were measured around smart utility meters operating at 868 MHz under laboratory-controlled conditions. The maximum 6-min averaged exposure recorded was 0.1 mWm⁻², which is less than 0.0024% of the corresponding 1998 ICNIRP general public reference level. Duty factors measured were less than 2.8%. This study found that the exposure contribution from Zigbee smart meter devices operating at 868 MHz is generally lower than, if not similar to, those operating at 2.4 GHz. © 2023 Crown copyright. Bioelectromagnetics published by Wiley Periodicals LLC on behalf of Bioelectromagnetics Society.